The overall objective of the project is to provide a HTS screening assay and paradigm to identify protein kinase inhibitors for Trypanosoma brucei that causes Human African trypanosomiasis (HAT), or sleeping sickness. HAT is endemic in sub-Saharan Africa, claiming the lives of about 30000 people every year and putting approximately 60 million people at risk of infection. HAT is not a priority for the pharmaceutical industry and thus the NIH and WHO categorized it as a neglected tropical disease (NTD). The need for new, effective chemotherapy is urgent. In order to identify new chemotherapy Dr Kojo Wilmot-Mensa (UGA) has used a chemical proteomic strategy using the kinase inhibitor, lapatinib, to identify kinases essential to the parasite. He identified three kinases that have additional genetic validation. In preliminary work these kinases were expressed, purified and enzyme assays established that demonstrated activity. Toward the goal of discovering new medicines for HAT, a multidisciplinary team has been established to develop an HTS assay and screening paradigm for these kinases. The team will employ a strategy to take advantage of both phenotypic and molecular approaches to drug discovery and therefore increase the probability of success through the 1) use of three T. brucei kinases, 2) full-length protein and substrates in assay formats that will capture 3) different molecular mechanisms of action, such as slow-dissociation, ATP noncompetitive inhibition and blocking kinase activation. The molecules identified will be clustered and representatives evaluated for activity in phenotypic parasitic assays. Our multidisciplinary approach merges the complementary skills of parasite biology and chemical proteomics (Dr Kojo Wilmot-Mensa, UGA), drug discovery and kinase enzymology (Dr David Swinney, non-profit Institute for Rare and Neglected Diseases Drug Discovery), kinase HTS assay development and drug discovery (Dr Chakk Ramesha, Medhus Bio) and protein expression and production (Dr David Chereau, 96 Proteins). At the end of the three-year funding period we expect to deliver a validated HTS assays and screening paradigm to deliver compounds with the quality to become medicines. We anticipated that assays developed through this initiative will be submitted to HTS facilities and we expect to use the results from HTS as the basis for seeking additional funding from appropriate entities for lead compound development.